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Updated: Jul 19, 2025
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The Isolation and Characterization of Low- and Normal- Density Neutrophils from Whole Blood
Published on: February 7, 2025
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Protocol for density gradient neutrophil isolation and flow cytometry-based characterization from human peripheral
Yi Kuang1, Upasana Parthasarathy1, Roberta Martinelli1
1Discovery Immunology, Merck & Co., Inc., Cambridge, MA 02141, USA.
STAR Protocols
|August 17, 2023
Summary
Isolating neutrophils, crucial immune cells, from blood is challenging. This protocol details a density gradient method for obtaining pure neutrophils for research on host immune responses.
Area of Science:
- Immunology
- Cell Biology
Background:
- Neutrophils are critical first responders in host defense against pathogens.
- Investigating fresh neutrophils presents significant challenges due to their sensitivity and rapid activation.
Purpose of the Study:
- To present a reliable protocol for isolating neutrophils from peripheral blood.
- To enable detailed morphological and immunophenotypic analysis of neutrophils.
Main Methods:
- Isolation of neutrophils using density gradient separation from peripheral blood.
- Morphological assessment via cytospin preparation.
- Immunophenotyping utilizing flow cytometry analysis.
Main Results:
- Successful isolation of neutrophils with high purity from healthy donors.
- Demonstration of protocol applicability to both healthy and diseased individuals.
Conclusions:
- This density gradient protocol provides a robust method for neutrophil isolation.
- The isolated neutrophils are suitable for downstream analyses, aiding research into immune responses.
